International Gluten Workshop, 11th; Beijing (China); 12-15 Aug 2012

1. Wheat quality improvement in
China, progress and prospects
Zhong-hu He12, Xianchun Xia1, Yan Zhang1, Yong Zhang1,
and Xinmin Chen1
1Crop Science Institute, CAAS
2Global Wheat Program, CIMMYT

2. Outlines
 Chinese wheat quality
 Progress in quality improvement
 Application of genomic technology
 Future prospects

3. Chinese wheat quality

4. Major cereal production in China, 2007-2009
Crop Area Yield Production %
Mha Kg/ha Mt World
Rice 29.5 6520 196 29
Maize 30.2 5327 161 20
Wheat 23.9 4703 112 17
Data Source: FAO Statistics, 2011

5. Wheat/maize interplanting
Wheat/maize rotation
Wheat/rice rotation
Chinese wheat production zones

7. Wheat consumption in China, 2009
Classification %
Food 77
Industrial use 9
Feed 9
Seed and storage loss 5
Total 100

8. Percentage of wheat foods in China
Food type %
Steamed bread including flat bread 45
Noodles and dumplings 40
Cookies and biscuits 8
Western bread 4
Others 3
Total 100

9. History of quality improvement
 Before 1984, focused on yield improvement and disease
resistance, little efforts on quality, white and hard kernel
means good quality
 1985-2000, establishment of quality labs and screening of
advanced lines and varieties, market demanding for better
processing quality
 2000-present, release of high quality variety, and
development and application of markers, market
demanding both for processing and nutritional qualities

10. Chinese wheat quality
 Broad variation for all major parameters, mixed
population, north with hard type, south with soft type
 Acceptable protein content, weak gluten strength and
poor extensibility
 Color needs improvement for traditional products
 Acceptable quality for manual processing, inferior
quality for mechanized production

11. 25
20
15
中国
China
10 加拿大
Canada
5
0
蛋白质
Protein 稳定时间
Stability 延伸性
Extensibility
Comparison between Chinese and Canada wheats

12. 100
80
软质
Soft
60
40 硬质
Hard
20
0
北方
North 南方
South
Distribution of kernel hardness in China

13. Genetic variation for bread-making quality

14. Genetic variation for noodle quality

15. Target
 Products: pan bread and white noodle for north China
 Traits: improvement of gluten strength and color rather
than protein content
 Region: Yellow and Huai Valley, 75% of production
 Impact in milling industry and farmer field, new
varieties with excellent quality and high yield
 Novelty in science, application of genomics and
proteomics, and molecular markers

16. Approach
 Integration of breeding, cereal chemistry, new
technology, and crop management
 National and international collaborations, CIMMYT,
Australia, USA, France, Japan, and UK….
 Training

17. Progress in quality improvement

18. Quality improvement
 Learn available technology for pan bread from other
countries, develop noodle testing methodology
 High yielding Chinese wheat crosses with germplasm from
CIMMYT, USA, and Australia, and then backcrosses with
Chinese wheat to ensure high yield and broad adaptation
 SDS sedimentation value, mixograph, and HMW-GS are
used as selection tool in early generations

19. 馒头制作方法
Noodle preparation and testing
Formula: flour extract rate 60%, water addition 35%, and salt 1%

20. New scoring system for Chinese white noodles
Character Chinese Japanese
Color 15 20
Appearance 10 15
Firmness 20 10
Viscoelasticity 30 25
Smoothness 15 15
Taste and flavor 10 15
Total 100 100
Zhang et al, 2005, Euphytica,141:113-120

22. Sensory scoring system for noodle quality
Name Origin Date
Parameter Excellent Very good Good Fair Poor Very poor Unacceptable
(10) (9) (8) (7) (6) (5) (4)
Color
(15)
Appearance
(10)
Firmness
(20)
Viscoelasticity
(30)
Smoothness
(15)
Taste & Flavor
(10)
Comprehensive
evaluation

24. Three factors for noodle quality
80.0
L* value of flour water slurry
 Medium-strong gluten quality 79.5
79.0
 High starch viscosity
78.5
 Bright color 78.0 r=-0.95
77.5
77.0
0.0 1.0 2.0 3.0 4.0 5.0
Flour colour grade
He et al, 2005, Cereal Chemistry, 82: 345-350
Zhang et al, 2005, Cereal Chemistry, 82: 633-638

25. Variety with outstanding noodle quality
 Excellent quality, higher yield than check variety
 Hebei: Jing 9428, Zhongmai 175
 Shandong: Yannong 15, Jimai 19, Jimai 20, PH82-2-2
 Henan: Yumai 34, Yumai 47, Yumai 49
 Introductions: Eradu, Gamenya, Sunstate

26. Variety with outstanding bread quality
 Excellent quality, yield close to check variety
 Accepted by farmers and milling industry
 Hebei: Zhongyou 9507, Gaocheng 8901, Jishi 02-1
 Shandong: Jinan 17, Jimai 20
 Henan: Yumai 34, Zhengmai 366
 Yumai 34 and Jimai 20 have dual purpose quality

27. Variety with improved bread-making quality

28. Application of genomic technology

29. Approach
 Breeding oriented approach, translate advances of
genomics into breeding programs, focus on
functional marker development and validation
 Functional markers can discriminate alleles of a
targeted gene, is an ideal marker for breeding
program
 Optimize available markers from other institutes

30. Example 1-yellow pigment
 Bright white color is preferred for Chinese noodle and
steamed bread, low yellow pigment is desirable
 Three QTLs at chr 7A, 7B, and 7D, are responsible for
yellow pigment
 Clone Psy 1 gene on chr 7A, 7B, and 7D
 Develop functional markers based on the gene allelic
variations
 Validate markers in Chinese wheat varieties

31. Cloned Psy genes on wheat chr 7A and 7B
Allele Coding Intron cDNA (bp) Deduced amino acids
seq (bp)
5’UTR ORF 3’UTR Residues Mass (kD)
PSY-A1 4177 bp 5 221 1284 303 428 47.8
PSY-B1 3313 bp 5 222 1263 156 421 47.0
1 2 3 4 5 6
Z MU32 636
ZMU 32636
1 5995
1 2 3 4 5 6
PSY-A1
1 4177
1 2 3 4 5 6
PSY-B1
1 3313
He et al, 2008, TAG, 116: 213-221

32. Allelic variants for the Psy-A1 gene on chr 7A
1 2 3 4 5 6
Psy-A1a
1 4177
1 2 3 4 5 6
Unt i t l e d
Psy-A1b
1 414 5
1 2 3 4 5 6
Psy-A1c
Unt itl e d
1 323 5
He et al, 2008, TAG, 116: 213-221

33. PCR amplification with YP7A
Varieties with high Varieties with low
yellow pigment yellow pigment
194 bp 231 bp
231 bp
194 bp

34. Validation of YP7A in Chinese varieties
Allele Accession number Mean (mg/kg) Range
Psy-A1a 130 1.80 a 0.62-3.42
Psy-A1b 87 1.35 b 0.35-2.88
Different letters indicate significant difference at P<0.05
He et al, 2008, TAG, 116: 213-221

35. Markers for color traits
 Yellow pigment: Psy-A1, Psy-B1, Psy-D1, TaZds-A1, and
TaZds-D1
 Polyphenol oxidase activity: PPO-A1 and PPO-D1
 Lipoxgenase activity: TaLox-B1
 Excellent understanding on color traits at molecular level
and powerful tool for breeders, genetic materials are
needed to develop markers for other locus

36. Example 2-LMW-GS
 Molecular marker development and application
 CE and MALDI-TOF-MS

37. Utilization of LMW-GS in breeding
 HMW-GS is well characterized and have been widely
used in breeding programs for more than 25 years
 LMW-GS are poorly characterized, and utilization in
breeding is not common, largely due to the unavailability
of simple and efficient method

38. Relationship between Glu-B3 protein alleles from
SDS-PAGE and gene haplotypes
Line Allele GluB3 GluB3 GluB3- GluB3- GluB3- GluB3- GluB3- GluB3-
-11 -12 13 14 15 21 22 23
Aroona-B3a a + +
Aroona b + +
Aroona-B3c c
Aroona-B3d d
Cheyenne e + +
Aroona-B3f f + +
Aroona-B3g g + +
Aroona-B3h h
Aroona-B3i i
Line Allele GluB3 GluB3 GluB3- GluB3- GluB3- GluB3- GluB3- GluB3- GluB3-
-31 -32 33 34 41 42 43 44 45
Aroona-B3a a +
Aroona b +
Aroona-B3c c + +
Aroona-B3d d + +
Cheyenne e +
Aroona-B3f f +
Aroona-B3g g +
Aroona-B3h h + +
Aroona-B3i i + +

39. Establishment of gene marker system and separation of
LMW-GS genes in Xiaoyan 54
3730 DNA analyzer
Zhang et al, 2011, TAG, 123: 1293-1305, adapted from Zhang Xiaofei, CAS

40. LMW-GS genes in Chinese core collections, CAS
17 16 17 16
Allelic variants of individual LMW-GS genes are conserved in sequences and polymorphic in length.

41. Markers for discrimination of Glu-B3 alleles
a b c d e f g h i
gluB3fg
812bp
gluB3g
853bp
gluB3h
1022bp
gluB3i
621bp

42. Molecular markers for Glu-A3 and Glu-B3
 Glu-A3: 7 markers for alleles a, b, c, d, e, f, and g
 Glu-B3: 9 markers for alleles a, b, c, d, e, f, g, h, and i
 Glu-D3: no marker is available due to tiny difference
in gene sequence, with minor effect on quality

43. Molecular marker validation
 More than 1000 varieties and advanced lines from
China, CIMMYT, and other 20 countries were tested
 Results from markers at Glu-A3 and Glu-B3, are well
consistent with SDS-PAGE
 Much simple to use

44. Multiplex PCR assay
 Screen 3-5 genes in one test
 Excellent accuracy
 Low cost
 Breeding oriented
Zhang et al, 2008, Plant Breeding, 127: 109-115

45. Application of markers
 Totally, 90 markers available in our lab
 Parental characterization and advanced lines confirmation
 Provision of very useful information for crossing program
with low cost, impossible by conventional method
 MAS operation in four breeding programs

46. Four lines in regional trials
Application of molecular markers in breeding

47. Glu-A3a
1 2 3 4 5 6 Glu-B3a
Glu-D3a Chinese Spring
2 Glu-B3h
5 CB037
HMW-GS
10
12 Glu-A3c
CB037-1
Glu-B3h Glu-B3h
Glu-A3a
Glu-A3c Glu-A3c CB037-2
Glu-B3d
Glu-B3g Glu-B3a
Glu-D3c
Ari124-3
Glu-D3a
Glu-D3c
Glu-B3g
Ari127-6
Rapid identification of LMW-GS alleles by capillary electrophoresis
Li et al, J. Cereal Sci, 2012, slide from Yan Yueming, Capital Normal University

48. Identification of Glu-B3 alleles by MALDI-TOF-MS
6. Chinese Spring, 7. Renan, 8. Insignia

49. MALDI-TOF-MS for LMW-GS
 Joint developed by the Capital Normal University and
Murdoch University
 A powerful and rapid method, 4-5 minutes per sample
 Little operational cost, breeding program can not
afford the equipment, need centralized service

50. Varieties recommended as standards for LMW-GS
Locus Subunit Standard cultivar
Glu-A3 Glu-A3a Neixiang 188, Chinese Spring
Glu-A3b Gabo, Pavon
Glu-A3c Pitic, Seri 82
Glu-A3d Nidera Baguette 10, Cappelle-Desprez
Glu-A3e Amadina, Marquis
Glu-A3f Kitanokaori, Renan
Glu-A3g Bluesky, Glenlea
Glu-B3 Glu-B3a Chinese Spring
Glu-B3b Renan, Gabo
Glu-B3b* Nanbu-komugi
Glu-B3c Insignia, Halberd
Glu-B3d/i Pepital, Norin 61
Glu-B3g Splendor, Cappelle-Desprez
Glu-B3g* Thesee, Aca 801
Glu-B3h Aca 303, Pavon
Glu-B3i* Heilo, Opata
Glu-B3j Grebee, Seri 82
Glu-D3 Glu-D3a Chinese Spring, Neixiang 188
Glu-D3b Gabo, Avocet
Glu-D3c Insignia, Cappelle-Desprez
Glu-D3c* Amadina, Heilo
Glu-D3f Ernest, Darius

51. Future prospects

52. Challenge 1- production
 Food security is China’s national policy, and
improvement of average yield is the only option
 Climate change, shortage of water, and more diseases
 Maize area increased 30%, wheat is less competitive than
maize in yield and price, can we maintain wheat area?
 Challenges are to combine high yield potential, disease
resistance, input use efficient, and excellent quality into
one variety

53. Challenge 2- quality
 Significant progress has been made in quality
improvement, still can not meet the needs of milling
industry
 Breeders give more efforts to yield improvement after
2005, due to the high price of average quality grain,
should we still breed for quality?
 Around 15 million tons wheat used for feed in 2012
 With small farmer’s size (0.5 ha), can we ensure quality
consistency?

54. Challenge 3-health food
 Health food is a hot subject, stop use of bleaching in
milling industry, genetic improvement of color
becomes more important
 Toxicity of Fusarium head scab could be a significant
threat, due to climate change and popularity of
wheat/maize rotation
 How should we prioritize Fe/Zn and other nutritional
elements? food diversity or genetic improvement?

55. Challenge 4- traditional products
 Significant progress has been made in improving white
noodle quality, but more efforts are needed to
understand other types of noodles, and dumplings
 More efforts to understand north style steamed bread
quality and Chinese flat bread….
 More efforts for soft wheat products

56. Opportunity 1-molecular marker
 Molecular markers will play an increasing and much
more significant role in varietal development within
next 5-10 years
 More and more functional markers will be available for
important traits, as progress made in genomics and
gene cloning
 Can we rapidly transfer the advances from sequencing
into breeding program?

57. Opportunity 2-GMO wheat
 Large investment on GMO crops including wheat
 Great potential for wheat improvement
 Low transformation efficiency: <1%
 Shortage of interest genes
 Declines of consumer’s acceptance

58. Opportunity 3- integrated approach
 Great expectations from new science in understanding
quality and provision of new tools
 Combination of conventional breeding, quality testing,
molecular markers, GMO, and other approaches
 International and national network

59. Conclusions
 Significant progress has been made in noodle testing
methodology and development of quality variety
 Comparative genomic was successfully used for gene
cloning, molecular markers development and
application, particularly for LMW-GS
 Integration of various disciplines, and combination of
traditional approaches and new sciences will help us to
meet the great challenges in the future

60. Collaborators
Liu Jianjun, Shandong AAS
Yan Yueming, Capital Normal University
D. W. Wang/A.M. Zhang, Chinese Academy of Science
R. J. Pena, CIMMYT
K. Quail/S. Huang, Former BRI
T. Ikeda/Yoshida, NARO
G. Branlard, INRA
W. J. Ma/R. Appels, Murdoch University
C. Morris, USDA-ARS

61. Acknowledgement
 Ministry of Agriculture
 Ministry of Science and Technology
 National Natural Science Foundation of China